Necrosis is a common pathologic feature of numerous diseases, but the molecular details regarding the execution of this pathway remain largely unknown. Our lab has produced significant insight into the mechanisms involved in necrotic cell death examining the anti-protease function of intracellular serpin proteins. Using the C. elegans model system, it has been shown that SRP-6 functions to block necrosis in response to multiple stress-inducing stimuli. More recent studies demonstrate that lack of SRP-6 results in impaired protein homeostasis, and that increased proteotoxicity results in necrotic cell death in either wild- type or srp-6 null animals. The long term objectives of this project are to dissect the relationship between proteotoxicity and necrotic cell death, and to identify specific genes involved in the regulation of these processes. The C. elegans model system will be used to examine the following specific aims: 1) to determine how the loss of SRP-6 perturbs protein homeostasis and 2) to determine how perturbed protein homeostasis results in necrotic cell death. In Aim 1 I will perform rescue experiments in srp-6 null animals to determine whether SRP-6 inhibitory activity is involved in the regulation of protein homeostasis. Additionally, I will employ an RNAi screen to identify specific peptidases as SRP-6 regulatory targets. In Aim 2, I will utilize both forward and reverse genetic approaches to identify specific genes that modulate the protein homeostasis and/or NCD pathways. The results of these studies will provide new insights into the mechanisms that regulate protein homeostasis and necrosis, facilitating the development of therapeutic options for treatment of necrosis-related diseases. PUBLIC HEALTH RELEVANCE: Necrosis is a common pathologic feature of many diseases including those associated with ischemic injury, chronic inflammation, and many gastrointestinal disorders. The goal of this research proposal is to determine the molecular mechanisms and to identify critical genes that contribute to necrotic cell death. Ultimately, these studies will provide the basis for development of highly specific treatments for patients with necrosis-associated disorders.